Effects of estradiol-17β and bisphenol A administered chronically to mice throughout pregnancy and lactation on the male pups＇ reproductive system

Aim： To assess the effect of estradiol-17β （E2） and bisphenol A （BPA） administered chronically by implanting a silicone tube throughout pregnancy and lactation on male pups＇ reproductive system in ICR mice. Methods： Female mice were implanted with a tube filled with 10 ng, 500 ng, 1 μg, or 10 μg of E2, or 100 μg or 5 mg of BPA, before mating. The tube was kept in the mice throughout pregnancy and lactation, until the pups had weaned at 4 weeks of age. During the period, E2 was released from the tube at 120 pg or 6, 12 or 120 ng/day, and BPA at 1.2 or 60 μg/day. Results： Most of the mice given 1 μg and 10 lag of E2 did not maintain their pregnancy. However, the other groups showed high rates of birth, more than 70%. At age of 4 weeks, the male pups were killed. Body weight and reproductive organ weights （testes, epididymides and accessory reproductive glands） in the treated groups did not differ from the control values, whereas the percentage of seminiferous tubules in the testis with mature spermatids was significantly lower in the groups given 10 ng and 500 ng of E2 and 5 mg of BPA than that in the control. Conclusion： Chronic exposure to E2 and BPA might disrupt spermatogenesis in male pups. （Asian JAndrol 2008 Mar; 10： 271-276）

Distribution of Chitinolytic Enzymes in the Organs and cDNA Cloning of Chitinase Isozymes from the Stomach of Two Species of Fish, Chub Mackerel (<i>Scomber japonicus</i>) and Silver Croaker (<i>Pennahia argentata</i>)

Chitinolytic activities were measured in two fish species having different feeding habits, chub mackerel (Scomber japonicus) and silver croaker (Pennahia argentata). Chitinase (an endo-type chitinolytic enzyme) activity was measured using pNP-(GlcNAc)n (n = 2, 3) as substrates;its level was significantly high in the stomachs of both species, as well as in the gills, intestine, pyloric appendage, testis, and liver of chub mackerel and in the spleen, kidney, pyloric appendage, ovaries, heart, and liver of silver croaker. β-N-Acetylhexosaminidase (an exo-type chitinolytic enzyme) activity was measured using pNP-(GlcNAc) as a substrate;it was detected at high levels in many parts apart from the digestive tracts of both species. The optimum pH for chitinase activity was 3.0 - 5.0 in the stomachs of both species, 4.0 in the liver of chub mackerel, and 4.0 and 8.0 in the kidney of silver croaker. Full-length cDNAs encoding two chitinase isozymes were obtained from the stomachs of the two fish species: SjChi-1 (1604 bp) and SjChi-2 (1512 bp) from chub mackerel and PaChi-1 (1630 bp) and PaChi-2 (1606 bp) from silver croaker. Expression analysis of these genes in the organs of the two species revealed strong expression of SjChi-1 in the stomach of chub mackerel and that of PaChi-1 andPaChi-2 in the stomach of silver croaker. The difference in the expression pattern of these genes is likely attributed to the difference in the feeding habits of the two fish species. Our results suggested the presence of novel chitinases in the two species.

Relationship between Nitrogen Atmospheric Deposition, Discharge and Concentration, and Monthly Change of Those in a River

The hypothesis that the product of discharge and concentration of nitrogen (N) in river water is equal to the atmospheric deposition was verified in the mountainous basin of the Tedori River in Japan. To verify this relationship, long-term data?are required to eliminate the effect of short-term variation of the N components. The basin has very high mountains, including Mount Hakusan (2702 maltitude), which is covered with deep snow in winter. Therefore, limited data were used for the estimation of the deposition of the entire basin by assuming a linear relationship of altitude. As a result, it was found that the estimated N concentration coincided well with observed concentrations at six sites—the Shiramine and Kuwajima (upper stream), Nakajima (lower stream) and Dainichi dam, Tedori dam and Senami sites (middle stream). The seasonal variation of N concentrations was low in the snowmelt period and high in autumn through to winter. This was not due to the larger discharge in snowmelt season as it was also found that N deposition was high in winter and low in spring, which indicated a clear relationship between N concentration and monthly atmospheric deposition including N storage in snow pack.

Suppression of postprandial blood glucose elevation by buckwheat(Fagpopyrum esculentum) albumin hydrolysate and identification of the peptide responsible to the function

The objective of this study is to evaluate the suppressive effect of buckwheat-albumin hydrolysate on postprandial hyperglycemia and identify the peptide responsible to the function. Buckwheat-albumin hydrolysate was prepared by using digestive enzymes and was orally administered to rats together with soluble starch. The blood was taken from the tail vein up to 90 min after oral administration to measure blood-glucose and plasma-insulin levels. The peptide with α-amylase inhibitory activity was purified from the buckwheathydrolysate by gel-filtration chromatography, α-amylase affinity chromatography, and high performance liquid chromatography(HPLC). The amino-acid sequence of the peptide was identified by a protein sequencer and was compared with that in the buckwheat-genome database. Buckwheat-albumin hydrolysate significantly suppressed the elevation of blood glucose level 15 min after starch administration. The amino-acid sequence of the peptide with α-amylase inhibitory activity was YVEPDCGNLGCCYHC in the parental protein of molecular mass 17.8 k Da and theoretical pI 4.77. The amino-acid sequence, molecular weight, and pI of the parental protein in buckwheat albumin were different from those of α-amylase inhibitor in wheat albumin. This study suggests that the novel α-amylase inhibitor identified in buckwheat albumin is a potential candidate for a functional food material to suppress postprandial blood glucose elevation.

Influence of Sowing Season and Host Crop Identity on the Community Structure of Arbuscular Mycorrhizal Fungi Colonizing Roots of Two Different Gramineous and Leguminous Crop Species

Introduction of cover crops may improve the diversity of arbuscular mycorrhizal fungi (AMF) in roots and soil under crop rotational systems;therefore, it is necessary to determine the potential for AMF communities to improve sustainable food production. We investigated the impact of cover crops, including wheat (Triticum aestivum L.), barley (Hordeum vulgare L.), pea (Pisum sativum L.), and hairy vetch (Vicia villosa Roth.), on the AMF communities in their roots in autumn and spring sowing seasons with PCR-DGGE analysis. Although all four cover crops impacted the AMF community structure in roots, the diversity of AMF communities was unchanged among crop type or sowing season. Redundancy analysis (RDA) demonstrated that AMF communities within crop type were significantly different. However, the AMF community structures were not influenced by growing season, suggesting that growth stage in crops may be more responsive to shaping AMF community structure in crop roots than host crop identity.

Distribution of Chitinolytic Enzymes in the Organs and cDNA Cloning of Chitinase Isozymes from the Liver of Golden Cuttlefish <i>Sepia esculenta</i>

The distribution of chitinolytic enzymes in eight organs of the golden cuttlefish Sepia esculenta was determined. Chitinase activity (activity of endo-type chitinolytic enzyme) was measured using pNP-(GlcNAc)n (n = 2, 3) as substrates, with high activity detected in the liver, posterior salivary gland, and stomach. β-N-acetylhexosaminidase (Hex) activity (activity of exo-type chitinolytic enzyme) was determined using pNP-(GlcNAc) as a substrate, and high activity was observed in six organs, including the liver, branchial heart, posterior salivary gland, and stomach. In addition, two chitin-binding proteins (CBP-A, CBP-B) were isolated from the liver using a chitin affinity column. Two full-length cDNAs (SeChi-1: 1484 bp;SeChi-2: 1748 bp) encoding chitinases were obtained from the liver of S. esculenta. SeChi-1 contained a 1377-bp open reading frame (ORF) encoding 459 amino acids, and SeChi-2 contained a 1656-bp ORF encoding 552 amino acids. Domain structures predicted from the deduced amino acid sequences of SeChi-1 and SeChi-2 (SeChi-1, SeChi-2) contained signal peptides, a GH Family 18 catalytic domain, one chitin binding domain (CBD) in SeChi-1, and two CBDs in SeChi-2. Proteome analysis revealed that 125 peptide residues of CBP-A were present in SeChi-1, and 116 peptide residues of CBP-B were present in SeChi-2. Organ expression analysis revealed that SeChi-1 and SeChi-2 were expressed only in the liver of S. esculenta. Phylogenetic analysis of SeChi-1, SeChi-2, and GH family 18 chitinases revealed that SeChi-2 belongs to a group of previously reported squid chitinases, while SeChi-1 does not belong to any previously reported group of mollusk chitinases.

Molecular Cloning and Phylogenetic Analysis of a Chitin Deacetylase Isolated from the Epidermis of the Red Snow Crab <i>Chionoecetes japonicas</i>

Chitin deacetylase (CDA;EC 3. 5. 1. 41) catalyzes the deacetylation of chitin. In this study, we successfully cloned and sequenced a chitin deacetylase gene from the red snow crab Chionoecetes japonicas. By using reverse transcription-polymerase chain reaction (RT-PCR) and 5' and 3' rapid amplification of cDNA ends, we obtained a 2141-bp amplicon containing a chitin deacetylase gene (CjCDA) from the epidermis of C. japonicas. The amplicon contains a 1575-bp open reading frame that is predicted to encode a 525-amino acid protein. The structure predicted from the deduced amino acid sequence included an N-terminal signal peptide, chitin-binding domain (CBD), low-density lipoprotein receptor class A domain (LDL-A), and catalytic domain. Comparative analysis of the deduced amino acid sequence of CjCDA revealed the highest homology (74%) to gastrolith protein 59 of Cherax quadricarinatus. We used RT-PCR to evaluate the expression of CjCDA in various tissues of C. japonicas, and we observed that CjCDA was expressed only in the epidermis. A phylogenetic analysis, using the amino acid sequences of CjCDA and other known chitin deacetylases, showed that CjCDA belonged to a group of crustacean chitin deacetylases. To our knowledge, this is the first study reporting the cDNA cloning of a chitin deacetylase from a crab.

Distribution of Chitinolytic Enzyme in the Organs and Molecular Cloning of a Novel Chitinase Gene from the Kidney of Marbled Rockfish <i>Sebastiscus marmoratus</i>

Actinopterygii express two types of chitinase (acidic fish chitinase-1 (AFCase-1) and acidic fish chitinase-2 (AFCase-2)) that are active at acidic pHs and involved in digestion in the stomach. We proposed the existence of a new fish chitinase that has a non-digestive function. In this study, we used Sebastiscus marmoratus, for which characteristics and cDNA cloning of chitinase isozymes (SmChi-1, SmChi-2) in the stomach have been reported. Initially, we examined the distribution of chitinase and β-N-acetylhexosaminidase (Hex) in the body and then we tried to clone novel chitinase cDNA from the kidney. Chitinase and Hex activities were measured using pNP-(GlcNAc)n, (n = 2, 3) and pNP-GlcNAc as substrates, respectively. Total RNA was extracted from the kidney. RT-PCR was performed to obtain chitinase cDNA fragments using reverse transcriptase with an oligo dT primer. The RACE method was used to obtain sequences of the upstream and downstream regions of cDNA. The full-length chitinase cDNA was determined using PrimeSTAR&reg;?Max DNA polymerase with proofreading activity. High chitinase activity was observed in the stomach, as previously reported. In addition, relatively high activity was observed in the liver, spleen, kidney, and heart. In contrast, Hex activity was detected in all organs. This result is consistent with the report that Hex is related to body-wide metabolism. Full-length cDNA (SmChi-3) of the novel chitinase was obtained from the kidney, which contained 1440 bp open reading frames. The domain structure of SmChi-3 was assumed to be similar to those of SmChi-1 and SmChi-2. SmChi-1 and SmChi-2 have a serine and glycine-rich linker region, which is characteristic of AMCase. In contrast, SmChi-3 contained no apparent sequence in the linker region. Phylogenetic analysis revealed the existence of a new chitinase group, which was named fish chitinase-3 (FCase-3) and differed from AFCase-1 and AFCase-2.

Potential Antimicrobial Effects of Gatifloxacin on Periodontopathic Bacteria in Vitro

The aim of this study was to identify the potential antibacterial effects of gatifloxacin on periodontal pathogens including Aggregatibacter actinomycetemcomitans, Porphyromonas gingi-valis, and Prevotella intermedia. The minimum inhibitory concentrations (MIC) of gatifloxacin and its bactericidal effects were investigated. Gatifloxacin inhibited the growth of all three kinds of periodontopathic bacteria tested in broth. The MIC value of 2.5 nM was found to be the most effective in inhibiting A. actinomycetemcomitans. An adenosine triphosphate biolumi-nescence assay revealed that gatifloxacin exhibited bactericidal effects on the tested bacteria in a time-dependent manner. The safety of gatifloxacin in mammalian cells was evaluated by assessing the viability of normal human dermal fibroblast (NHDF) cells treated with gatifloxacin. Almost all NHDF cells survived after 2-d culture, while 81% of the cells survived after 4-d culture when treated with 1.0 × 103 nM gatifloxacin. These results indicate that gatifloxacin is a possible drug for local administration to prevent periodontal infection.

Effects of acquiring FSC forest management certification for Japanese enterprises using SmartWood Audits

We explore the organizational, environmental, and economic effects of sustainable forest management （FM） certification by Smart- Wood （SW）, one of the certification bodies accredited by the Forest Stewardship Council （FSC） to carry out certification in Japan. We closely investigated three enterprises： the Yusuhara Forest Owners＇ Cooperative, Yamanashi Prefectural Forest, and Ryujin-mura Forest Owners＇ Coopera- tive. Interviews with representatives of these entities provided crucial empirical information regarding the influence of certification on envi- ronmental assessment, rare-species protection, landscape-management planning, management objectives, and forest supervision. Attainment of FM status improved environmental awareness and engendered positive changes in assessing and managing forests and other natural resources but simultaneously imposed heavier economic costs.

Discovery of the strong antioxidant selenoneine in tuna and selenium redox metabolism

A novel selenium-containing compound,selenoneine,has been isolated as the major form of organic selenium in the blood and tissues of tuna.Selenoneine harbors a selenium atom in the imidazole ring,2-selenylNα,Nα,Nα-trimethyl-L-histidine,and is a selenium analog of ergothioneine.This selenium compound has strong antioxidant capacity and binds to heme proteins,such as hemoglobin and myoglobin,to protect them from iron auto-oxidation,and it reacts with radicals and methylmercury(MeHg) .The organic cations/carnitine transporter OCTN1 transports selenoneine and MeHg,regulates Se-enhanced antioxidant activity,and decreases MeHg toxicity.Thus,the dietary intake of selenoneine,by consuming fish,might decrease the formation of reactive oxygen radicals that could oxidize nucleotides in DNA,and thereby inhibit carcinogenesis,chronic diseases,and aging.

Quantitative Analysis of the Rate of Geochemical Weathering of Sulfur from Sedimentary Rocks Using Atmospheric Deposition, Concentration and River Discharge Data

Quantitative analysis of the rate of geochemical weathering of sulfur (S) from sedimentary rocks (GeoS) was conducted using concentration (Cs) and discharge (Qs) data from the Tedori River and atmospheric deposition (AtdepS) in the basin. First, S fluxes were calculated using 16 years of Cs and Qs data. The annual average discharge of S (TotalS) was estimated at 8597 ton·year-1 (117.3 kg·ha-1·year-1). Of this, 1331 ton·year-1 was AtdepS (18.2 kg·ha-1·year-1) and another 7266 ton·year-1 was GeoS (99.1 kg·ha-1·year-1). Monthly changes in TotalS were investigated, which showed that GeoS was highest in summer, because of the air temperature, while AtdepS peaked in winter because of seasonal wind. Using Qs and AtdepS corrected for altitude, TotalS, AtdepS and GeoS were estimated at six sites, and among these sites we found that the TotalS per unit area values were random, depending on the site characteristics. In particular, the discharge from the Kuwajima site was remarkably high suggesting that the sedimentary rocks at this site had higher pyrite content than at the other sites. Finally, we also assessed the relationship between the characteristics of sedimentary rocks and GeoS in a range of rivers in the Hokuriku Region, and found that there was a close relationship between concentrations of SO42- greater than 10 mg·l-1 and sedimentary rocks containing the pyrite group. In addition, we estimated that the influence of GeoS was present when the concentration of SO42- in river water was greater than 2 - 3 mg·l-1 in the Hokuriku region.

Long-Term Assessment of Nitrogen Pollution Load Potential for Groundwater by Mass Balance Analysis in the Tedori River Alluvial Fan Area, Japan

To evaluate the nitrogen pollution load in an aquifer, a water and nitrogen balance analysis was conducted over a thirty-five year period at five yearly intervals. First, we established a two-horizon model comprising a channel/soil horizon, and an aquifer horizon, with exchange of water between the aquifer and river. The nitrogen balance was estimated from the product of nitrogen concentration and water flow obtained from the water balance analysis. The aquifer nitrogen balance results were as follows: 1) In the aquifer horizon, the total nitrogen pollution load potential (NPLP) peaked in the period 1981-1990 at 1800 t·yr-1;following this the NPLP rapidly decreased to about 600 t·yr-1 in the period 2006-2010. The largest NPLP input component of 1000 t·yr-1 in the period 1976-1990 was from farmland. Subsequently, farmland NPLP decreased to only 400 t·yr-1 between 2006 and 2010. The second largest input component, 600 t·yr-1, was effluent from wastewater treatment works (WWTWs) in the period 1986-1990;this also decreased markedly to about 100 t·yr-1 between 2006 and 2010;2) The difference between input and output in the aquifer horizon, used as an index of groundwater pollution, peaked in the period 1986-1990 at about 1200 t·yr-1. This gradually decreased to about 200 t·yr-1 by 2006-2010. 3) The temporal change in NPLP coincided with the nitrogen concentration of the rivers in the study area. In addition, nitrogen concentrations in two test wells were 1.0 mg·l-1 at a depth of 150 m and only 0.25 mg·l-1 at 50 m, suggesting gradual percolation of the nitrogen polluted water deeper in the aquifer.

<i>In Vitro</i>Culture of Shell Gland Epithelial Cells in Japanese Quail (<i>Coturnix coturnix japonica</i>)

To investigate the secretion of protoporphyrin IX (PpIX), superficial eggshell pigment, from shell gland cells of Japanese quail, the epithelial cells of the gland were collected and isolated for cultivation in vitro. An analysis of a peak for PpIX in the cells was performed using a fluorescence microplate reader. The measurement showed that PpIX has a peak of excitation wavelength at 410 nm and emission wavelength at 606 nm in the culture medium (HamF12 + 4% HCl). Volumes of PpIX in the medium after 4 hour culture of the cells were measured with a microplate reader using filter set of excitation wavelength 400/30nm and emission wavelength 620/40nm. However the cells did not secrete significantly PpIX during 4 hour incubation in this culture system, addition of quail plasma to the medium resulted in significantly higher secretion. A cultivation system in this study is able to use for the study on the mechanism of the secretion of eggshell pigment, PpIX from Japanese quail shell gland epithelial cells.

Calcium Participates in Secretion of Porphyrin from Shell Gland Epithelial Cells of Japanese Quail (<i>Coturnix coturnix japonica</i>)

To investigate factors involved in the secretion of protoporphyrin IX (PpIX), a superficial eggshell pigment, from shell gland epithelial cells of Japanese quail, we cultured cells in Ham’s F12 medium with calcium chloride and quail plasma. The addition of hormones (prostaglandin F2α, progesterone, estradiol-17β) to the medium did not change the PpIX concentration in the culture supernatant, but changing the calcium chloride (CaCl2) concentration did: a lower concentration of CaCl2 led to a higher PpIX concentration;0 mM CaCl2 enhanced the secretion of PpIX from epithelial cells prepared at 5 or 7 mM CaCl2. The result suggests that a drop in concentration of CaCl2 mimics the end of shell calcification and stimulates rapid secretion of PpIX in vivo. Bovine serum albumin was almost as effective as quail plasma for PpIX secretion in culture, and would facilitate further study of the mechanism of PpIX secretion.

Relevance of Visual, Auditory, and Olfactory Cues in Pet Dogs’ Awareness of Humans

Many studies have looked at how dogs respond to human communicative information. Here, we examined which human communicative factors were important in influencing dogs’ responses. Eleven healthy pet dogs with no apparent aggressive behaviour toward people were recruited. Five sensory conditions (all cues presented;either a visual, an auditory, or an olfactory cue presented;no cues presented) were provided three times randomly to each dog during the tests. All tests were video recorded, and both the dogs’ behaviour and time taken to reach the person when she presented each of the sensory cue conditions were observed. Total rates of reaching the person were as follows: 97.0% (all cues), 87.9% (auditory cues), 84.4% (visual cues), 84.4% (olfactory cues), and 69.7% (no cues). The time taken for the dog to notice the person in the box and then obtain a reward from her differed among the five conditions: all cues (6.00 ± 0.32 s) and visual cues (6.02 ± 0.91 s) were significantly faster than auditory cues (18.56 ± 9.57 s) and no cues (26.55 ± 11.72 s). Thus the type of information input was important in recognition of the person by the dogs and influenced the dogs’ response times;visual cues appeared advantageous in confirming the person’s presence.

A New Species of the Genus Rhabdophis Fitzinger,1843(Squamata:Colubridae)in Southwestern Sichuan,China

The genus Rhabdophis is a group of widely distributed snakes with more than 20 species.Recent field surveys uncovered a species in southwestern China,which has long been considered as R.pentasupralabialis.Combined molecular and morphological analyses revealed it as a new species Rhabdophis chiwen sp.nov.Based on 12 specimens,this new species is distinguished by the following characters:1)dorsal body saddlebrown,dorsal scales typically with black margins forming spots and stripes,the margin of the outer row forming two faint dorsolateral black cross-bars alongside body;2)ventral scales 151–159,the outer margin of ventral scales and several lateral rows of dorsal scales forming ventrolateral longitudinal brownish-red coloration,with faint black spots in the middle of ventral scales;3)a black oblique stripe present below eyes,often with a black spot between the 2 nd and 3 rd supralabial and a black stripe on the 5 th supralabial;4)eyes dark khaki,pupils black;5)infralabials usually 7,the first four in contact with anterior chin-shields;6)temporal scales 1+1;7)dorsal scales in 15 rows,feebly keeled except the outer 1–2 rows;8)anal scale divided;subcaudals 45–59;9)preocular 1 and postoculars 3(occasionally 2);10)body medium-sized(snout-vent length:adult males 404–431 mm,adult females 409–476 mm);11)tail moderate(tail length/total length in adult males 0.205–0.238,in adult females0.172–0.193).With the discovery of this new species,the total number of species in genus Rhabdophis is 28 with 12 th species known to occur in China.

cDNA Cloning of Paramyosin from Several Kinds of Squid Mantle Muscle

Paramyosin is a rod-shaped muscle protein found exclusively in invertebrates, with α-helices coiled around each other to form a coiled-coil structure. Marine organisms in which the primary structure of paramyosin has been determined are mollusks, including abalone (Haliotis discus), mussels (Mytilus galloprovincialis), octopus (Octopus bimaculoides), and oyster (Crassostrea gigas). In contrast, the primary structure of squid paramyosin, which is of particular interest, has yet to be reported. In the present study, cDNA cloning of paramyosins from four squid species, the neon flying squid (Ommastrephes bartramii), the Humboldt squid (Dosidicus gigas), the golden cuttlefish (Sepia esculenta), and the clawed armhook squid (Gonatus onyx), was performed to determine the following: the 2605-bp O. bartramii paramyosin gene containing a 2574-bp open reading frame (ORF), the 2691-bp D. gigas paramyosin gene containing a 2640-bp ORF, the 2631-bp S. esculenta paramyosin gene containing a 2574 ORF, and the 2609-bp G. onyx paramyosin gene containing a 2574-bp ORF. The primary structure of the four squid paramyosins was found to contain heptad repeats and an ACD (assembly competence domain), which are characteristic of a coiled coil. A phylogenetic analysis was performed with paramyosin sequences from species including the four squid species examined in this study, the results of which indicated that the four squid paramyosins form a group independent from the paramyosins of other species, to which octopus paramyosins are closest.

An Anionic Polymer Incorporating Low Amounts of Hydrophobic Residues Is a Multifunctional Surfactant. Part 1: Emulsifying, Thickening,Moisture-Absorption and Moisture-Retention Abilities of a FattyAcid-Containing Anionic Polysaccharide

We have been studying the function and structure of fatty acid-containing extracellular polysaccharides (FACEPS) produced by bacteria belonging to the genus Rhodococcus. In this study, we examined the relationships between the structure and emulsifying, thickening, moisture-absorption, and moisture-retention capabilities of rhodococcal FACEPS using S-2 EPS produced by R. rhodochrous strain S-2. We prepared chemically deacylated S-2 EPS (DeAcyl S-2 EPS) and palmitoylated DeAcyl S-2 EPS (ReAcyl S-2 EPS), and compared them with native S-2 EPS. All of the properties were attenuated and recovered by deacylation and reacylation of S-2 EPS, respectively. These results suggest that the fatty acid moiety of rhodococcal FACEPS is involved in such functional properties. We also showed that palmitoylation improved the emulsifying, moisture-ab-sorption, and moisture-retention abilities of other acidic polysaccharides that are commercially available. These results suggest that the acidity of the polysaccharide backbone is at least partly responsible for the observed functionality of fatty acid-containing polysaccharides. To our knowledge, this is the first report on multifunctional property of an anionic polymer incorporating low amounts of hydrophobic residues. The present findings could be useful for the creation of new multifunctional surfactants from renewable raw materials for use in various industries, e.g., in cosmetics.

26S proteasome inhibitors inhibit dexamethasone-dependent increase of tyrosine aminotransferase and tryptophan 2,3-dioxygenase mRNA levels in primary cultured rat hepatocytes

Dexamethasone (Dex), a ligand for transcriptional enhancement of tyrosine aminotransferase (TAT) and tryptophan 2,3-dioxygenase (TO) genes, (100 nM) maximally increased these mRNA levels at 12 h and 7 h in primary cultured rat hepatocytes and the nuclear fraction, respectively. Lactacystin (5 μM) and epoxomicin (0.5 μM), 26S proteasome inhibitors, significantly suppressed the Dex-dependent maximum increase of TAT and TO mRNA levels in the cells and the nuclear fraction. Electrophoretic mobility shift assay demonstrated that lactacystin did not affect binding of glucocorticoid receptor to glucocorticoid responsive element. Furthermore, lactacystin did not affect the activation of GRE luciferase reporter by Dex transfected to the cells. The results demonstrate that 26S proteasome is positively involved in the Dex-dependent increase of TAT and TO mRNA levels in the cells and suggest that the mechanism of action of 26S proteasome may be degradation of some RNase(s), which breaks down TAT and TO mRNAs.